1.Establishment and characterization of a human renal carcinoma cell line RCC-9863
Pengfei WANG ; Changhong SHI ; Chen SHAO
Chinese Journal of Urology 2001;0(06):-
Objective To establish a new renal cancer cell line so as to provide an useful tool for the basic research of renal cancer. Methods A renal tumor specimen sized 1.0 cm?1.0 cm was excised and inoculated subcutaneously into the right posterior leg of three nude mice.After three passages in the nude mouse,the tumor tissue was cultured in vitro. Following the reference, the identification of cell line were fulfilled. Results Two of the transplanted tumors grew in the nude mice and then part of the tumor was cut and inoculated again into nude mouse for another two generations. The tissue cultured in vitro was followed and finally a stable growth renal carcinoma cell line was established without change of morphological structure and differentiation. The mode rage of chromosome has been 62~68 (75%) and the cycle analysis showed G1 63.8%, G2 11.4% and S1 24.8%.The doubling time has been 37.7 hours.The tumor cell can secrete IL 6 continuously. Conclusions The renal carcinoma cell line RCC 9863 was identical to the primary cancer cell in biological characteristics.It has been cultured in vitro continually for more than one year with its characteristics remained.So,RCC 9863 is considered to be a stable cell line.
2.Analysis of Influential Factors of ADR Induced by Reduning Injection in Pediatric Patients of Our Hospital
Changcheng SHI ; Pengfei ZHANG ; Tianshou QI
China Pharmacy 2016;27(26):3636-3639,3640
OBJECTIVE:To observe the characteristics and influential factors of aderverse drug reactionl(ADR)induced by Re-duning injection. METHODS:The clinical information of pediatric patients receiving Reduning injection were collected from pediatric department of our hospital during Nov. 2014-Oct. 2015. Using ADR as outcome indicators,case control in the queue design was con-ducted. The influential factors of ADR induced by Reduning injection were analyzed by using binary univariate and multivariate Logis-tic regression analysis,Cox regression fitted 1∶3 matching Logistic regression analysis. RESULTS:Of 9 187 cases,the average age of children was(4.21±2.63)year-old;there were 23 ADR cases,with the incidence of 0.25%. There were 21 cases of rash(9 cas-es of combined pruritus)and 2 severe ADR cases(including 1 case of twitching limbs,blue complexion,peripheral cold,high fe-ver,leg pain and 1 case of limbs cold and high fever);23 cases of ADR symptoms finally disappeared after treated. Based on drug dose and usage of low concentrations and low drip rate in our hospital,regression analysis showed that age,sex,drug dose,solvent amount,concentration,drip rate,source of children,drug combination,the group number of daily continuous intravenous infusion, infusion order,the days of continuous medication and other factors were not statistically significant (P>0.05);the incidence of ADR induced by Reduning injection was 0.11% in the group of 1-3 years old children and was 0.31% in the group of 3-16 years old children. There was no statistical significance between 2 groups(P>0.05). CONCLUSIONS:The reasons for ADR induced by Re-duning injection might be the drug itself,the wrong methods,etc. Reasonable drug dosage,low concentration,low drip rate,using sophisticated filters infusion device and changing infusion device while dressing can reduce the occurrence of ADR.
3.Chemical constituents of Abrus mollis
Jing WEN ; Haiming SHI ; Pengfei TU
Chinese Traditional and Herbal Drugs 1994;0(05):-
Objective To study the constituents of Abrus mollis.Methods The constituents were isolated by chromatographic methods,their structures were elucidated by spectroscopic evidences.Results Eleven compounds were purified and their structures were identified,they were identified as ?-sitosterol(Ⅰ),stigmasterol(Ⅱ),nonacosanyl caffeate(Ⅲ),daucosterol(Ⅳ),betulinic acid(Ⅴ),vanillic acid(Ⅵ),inositol methyl ether(Ⅶ),sucrose(Ⅷ),soyasaponin Ⅰ(Ⅸ),kaikasaponin Ⅲ(Ⅹ),and dehydrosoyasaponin Ⅰ(Ⅺ).Conclusion Compounds Ⅰ,Ⅲ,Ⅳ,Ⅵ,and Ⅶ-Ⅺ are isolated from this plant for the first time.
4.Orthotopical transplantation of human renal carcinoma tissue into nude mice and the establishment of a high metastatic cell line MRCC
Pengfei WANG ; Changhong SHI ; Chen SHAO
Chinese Journal of Urology 1994;0(02):-
Objective To establish a SOI model of human renal carcinoma and a high metastatic cell subline. Methods A human renal cell line RCC-9863 has been established by inoculating a human renal tumor tissue into nude mice s.c..When RCC-9863 passaged for 20 times,the tissue from the same xenotransplant tumor were used to construct SOI model.Cultured the metastatic tissue in vitro,the tumor cell suspension was then injected orthotopically.The metastatic tissue obtained underwent the same procedure again.At last,the metastatic tumor was cultured in vitro and cloned. Results 15 days later, a tumor mass sized 1.7 cm?0.6 cm in the nude mouse's renal parenchyma was grown which lobulated,rude,and with multiply blood vessels and 55 days later the mouse became moribund and metastases in the lungs were formed.The transplanted renal tumor in the SOI model grew fast and invasively and metastasized to lungs,lymphatic node and liver.A subline,MRCC,with metastatic ability to the lung was selected.Compared with RCC-9863,MRCC has the characteristics of shorter multiply time and higher agar clone forming rate(P
5.Ultra High Performance Liquid Chromatography Coupled with Q Exactive Hybrid Quadrupole-orbitrap Mass Spectrometry for Serumal Metabonomics Study of Drug-induced Liver Injury in Patients
Zhuoling AN ; Chen SHI ; Rui ZHAO ; Pengfei LI ; Lihong LIU
Chinese Journal of Analytical Chemistry 2015;(9):1408-1414
Ultra high performance liquid chromatography coupled with a Q Exactive hybrid quadrupole-orbitrap mass spectrometry ( UPLC-MS/MS) and multivariate statistical analysis was applied to metabonomic
study of drug-induced liver injury. This investigation focused on the liver injury in clinic which was induced by antitumor drug, traditional Chinese medicine, statins and antibiotics. After precipitated by cold acetonitrile, the serum samples were separated on a HSS T3-C18 column using water (containning 0. 01% formic acid) and acetonitrile as mobile phase by gradient elution. Based on metabonomic profiles analysis of the orthogonal projections to latent structures discriminant analysis ( OPLS-DA ) and partial least squares-discriminant analysis ( PLS-DA) , 114 endogenous metabolites had been found to have significant differences between drug-induced liver injury patients and healthy volunteers, 38 endogenous metabolites have significant difference (p<0. 001). According to the results of the OPLS-DA, MS/MS data and human metabolome database ( HMDB) , phenylalanine and dimethylguanosine had been identified as potential biomarkers related to drug-induced liver injury for early discovery and diagnosis.
6.Ultrasonic Extraction Process Research and Mathematic Simulation of Tyrosol in Rhodiolae Crenulatae Radix et Rhizoma
Yufang HUANG ; Xiaofeng SHI ; Juli YUAN ; Pengfei XIA
Chinese Journal of Information on Traditional Chinese Medicine 2015;22(11):89-91
Objective To study the ultrasonic extraction technology of tyrosol in Rhodiolae Crenulatae Radix et Rhizoma;To mathematically simulate the extraction process.Methods The content of tyrosol was set as index;HPLC was used;the ultrasonic extraction process of tyrosol in Rhodiolae Crenulatae Radix et Rhizoma was optimized by an orhogonal experiment and mathematic simulation.Results The optimum ultrasonic extraction conditions were as follows:80%ethanol was used as extraction solvent, the ratio of material to liquid was 1:35 (g:mL), with the extraction time of 30 min and ultrasonic power of 240 W. In these conditions, the extracting rate of tyrosol was 0.150 1%. Compared with the heating reflux method, the extraction time should be shortened by 66.7% and the extracting rate should be increased by 12%.Conclusion The extraction method is simple and the extraction rate of effective components is high. Mathematical simulation values based on ultrasonic extraction are consistent with experiment values.
7.Using the External Quality Assessment Return Target Value Obtained from Clinical Biochemical Analyzer to Evaluate the Accuracy After the Regression Calibration of Dry-type Biochemical Analyzer
Guanghua SHI ; Huiying JIANG ; Pengfei WANG ; Qing ZHANG ; Lianyi LIU
Journal of Modern Laboratory Medicine 2014;(6):159-161
Objective Using Hitachi 7170 external quality assessment return target value to evaluate the accuracy of 10 items after Regression calibration of the Vistros 350 dry-type Biochemical Analyzer.Methods The same quality control samples were separately tested on two instruments,and results were reported to the clinical National Center for Clinical Laborato-ries.Substituted the external quality assessment return target value result from the National Center for Clinical Laboratories by using Vitros350 into regression calibration equation,then the getting data were compared with the external quality assess-ment return target value obtained from Hitachi 7170,and the deviation analysis was processed.The total error range from the America Clinical Laboratory Amended Bill was used as the standard.For the results within the reference range,error less than 1/2CLIA’88 total error,taken as the comparable judging standard,as it satisfied the requirement.For the results out off the reference range,error less than CLIA’88 total error,those still satisfied the requirement.For those items not meet the requirements,it must to do the regression calibration for Vitros350,using Hitachi 7170 as the standard instrument.Results The deviations of 7 items were all less than 1/2CLIA’88 allowed total error,with LDH was 0.16~-9.89,CK 2.92~6.25, ALT -4.64~-8.07,TBIL 0.08~2.67,TP -0.37~4.41,ALB 2.74~4.77 and URIC 1.04~3.0 respectively,and did not need re-calibration.For GLU and CREA,only one out of the reference range sample,the error range was >1/2CLIA’ 88,but
8.The application of implantable venous access ports in management of postoperative breast cancer patients
Zhimin LIU ; Yuan ZHONG ; Xueqing JIANG ; Pengfei SHI ; Yang LI
Chinese Journal of Endocrine Surgery 2016;10(2):148-151
Objective To investigate and analyze the experience of planting and maintaining implantable venous access ports(IVAP) in management of post-operative breast cancer patients. Methods Breast cancer pa-tients receiving IVAP after surgery from Mar. 2011 to Jun. 2014 were retrospectively analyzed. The relative com-plications were documented and summarized during implanting operation. Results 468 patients received IVAP, among whom 451 patients underwent piercing implantation via right internal jugular vein, 15 patients underwent piercing implantation via right subclavian vein, and 2 patients underwent piercing implantation via left internal jugular vein. The mean cathe tering leng th was 12.8 cm for patients receiving IVAP via right internal jugular vein, ranging from 12 to 15 cm. Thereinto, 30(6.4%) patients experienced shot-term complications including 16 cases of puncture difficulty, 5 cases of accidental arterial puncture, 2 cases of extravasation, 2 cases of blood aspiration dif-ficulty and 5 cases of arrhythmia. Three cases had long-term complications as the following:one case of catheter-re-lated infection, one case of catheter lost, and one case of incision rupture. Conclusions IVAP is a safe and effec-tive intravenous infusion device. It is crucial to choose individualized implanting access and length by professional surgical team.
9.Establishment of TaqManˉBHQ probe realˉtime fluorescence RTˉPCR for quantitative detection of Soul virus
Chuncheng LIU ; Pengfei YANG ; Qingli YAN ; Yujun SHI
International Journal of Laboratory Medicine 2014;(24):3332-3333
Objective To establish a rapid method of real-time fluorescence RT-PCR to quantify Soul virus.Methods The pro-fessional software was adopted to design the primer and the TaqMan-BHQ probe.With artificially synthesized L gene segment as the template of Soul virus,the real-time RT-PCR for detecting Soul virus was researched.Results The Ct value of templates had a good linear relationship with the log value of the template diluted concentration.The standard curve was Y =-3.607X +41.84, r2 =0.998,the PCR amplification efficiency was 108.1%,its lowest detection limit was 53.2 copies/μL.Conclusion Applying the real-time fluorescence RT-PCR by the TaqMan-BHQ probe for detecting nucleic acid of Seoul virus has the characteristics of short time-consuming and high sensitivity.
10.Comparison of osteopontin and mineralization culture medium in the induction of dental pulp stem cells into osteoblasts
Xin SHI ; Pengfei ZHANG ; Mengtong YUAN ; Mingyue LIU ; Weiping HU
Journal of Practical Stomatology 2015;(1):11-14
Objective:To compare the osteogenic differentiation of dental pulp stem cells(DPSCs)induced by osteopontin(OPN)and mineralizing culture medium(MCM).Methods:DPSCs were cultured with OPN(OPN group)and MCM(MCM group)respectively. The morphology of the DPSCs were observed under inverted microscope.The mineralize nodules were observed by alizarin red staining. RT-RCR was used to detect the mRNA expression of bone sialoprotein (BSP),Runt-related transcription factor 2(Runx-2),osteocal-cin(OCN)and collagen-1(Col-1).Results:Similar number of mineralized nodules was found in the 2 groups(P >0.05)after 28 day culture.The mRNA expression level of BSP gene in OPN group was higher than that in MCMgroup(0.864 ±0.112 and 0.514 ±0.068, P <0.05),while the expression level of Runx-2 gene in OPN group is lower than that in MCMgroup(0.186 ±0.017 and 0.324 ±0. 058,P <0.05).The expression level of Col-1 and OCN genes in both groups were similar(P >0.05).Conclusion:The capabilities of OPN and MCMin inducing osteogenic differentiation of DPSCs are similar.